Two Channel Chemistry and Imaging on the MiSeq i100 Series

Background

The MiSeq i100 Series uses two-color chemistry, which allows for rapid imaging of the flow cell using blue and green fluorescent excitation channels at every sequencing cycle. The MiSeq i100 Series is powered by XLEAP-SBS chemistry, a faster, higher quality, and more robust update to Illumina's sequencing by synthesis (SBS) chemistry.

Chemistry

In two-channel chemistry, the base calls are assigned by the fluorescent excitation-emission spectra that are designated by that base. The MiSeq i100 Series uses the following designations:

• Clusters with intensity in green only: T

• Clusters with intensity in blue only: C

• Clusters with intensity in both channels: A

• Clusters with no intensity/dark: G

  

CMOS Flow Cell

The MiSeq i100 Series uses a patterned flow cell with nanowells fabricated onto a CMOS chip.

Imaging Tiles

5M: 1 lane x 9 swaths x 2 tiles = total of 18 tiles imaged.

25M: 1 lane x 9 swaths x 10 tiles = total of 90 tiles imaged.

50M: 2 lanes x 9 swaths x 10 tiles = total of 180 tiles imaged.

100M: 4 lanes x 9 swaths x 10 tiles = total of 360 tiles imaged.

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